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Biosafety
Level 2 (for work with Wangiella
dermatitidis or other class II pathogens)
Biosafety Level
2 is similar to Level 1 and is suitable for work involving
agents of moderate potential hazard to personnel and the
environment. It
differs in that (1) laboratory personnel must have specific
training in handling pathogenic agents and are directed by
competent scientists, (2) access to the laboratory must be
limited when work is being conducted, (3) extreme precautions
must be taken with contaminated sharp items, and (4) certain
procedures in which infectious aerosols or splashes may be
created must be conducted in biological safety cabinets or
other physical containment equipment.
The following standard and special practices, safety
equipment, and facilities apply to agents assigned to
Biosafety Level 2:
A.
Standard
Microbiological Practices
1.
Access to the laboratory is limited and restricted at
the discretion of PJS when experiments are in progress.
2.
Experimenters must wash their hands after they handle
viable materials and animals, after removing gloves, and
before leaving the laboratory.
3.
Eating, drinking, smoking, handling contact lenses, and
applying cosmetics are not permitted in the laboratory work
areas. Persons
who wear contact lenses in laboratories should also wear
goggles or a face shield.
Food is to be stored outside the work area in the
refrigerator in ESB 109 designated for this purpose only.
4.
Mouth pipetting is prohibited; mechanical pipetting
devices must be used.
5.
All procedures are must be performed carefully to
minimize the creation of splashes or aerosols.
6.
Work surfaces must be decontaminated at least once a
day and after any spill of viable material.
7.
All cultures, stocks, and other regulated wastes must
be decontaminated before disposal by approved decontamination
by autoclaving. Materials to be decontaminated outside of the immediate
laboratory must be placed in a durable, leakproof container
and closed for transport from the laboratory.
Materials to be decontaminated off-site from the
laboratory must be packaged in accordance with applicable
local, state, and federal regulations, before removal from the
facility.
8.
An insect and rodent control program is in effect, and
each investigator must monitor the laboratory for insects and
rodents.
B.
Special Practices
1.
Access to the laboratory is limited or restricted by
PJS when work with infectious agents is in progress.
In general, persons who are at increased risk of
acquiring infection or for whom infection may be unusually
hazardous are not allowed in the laboratory or animal rooms.
For example, persons who are immunocompromised or
immunosuppressed may be at increased risk of acquiring
infections. PJS
has the final responsibility for assessing each circumstance
and determining who may enter or work in the laboratory.
2.
PJS will establish all policies and procedures whereby
only persons who have been advised of the potential hazard and
meet specific entry requirements (e.g., immunization) enter
the laboratory or animal rooms.
3.
When infectious agents are in use in the laboratory,
which require special provisions for entry (e.g.,
immunization) a hazard warning sign incorporating the
universal biohazard symbol will be posted on the access door
to the laboratory work area.
The hazard warning sign will identify the infectious
agent, list the name and telephone number of the laboratory
director or other responsible person(s), and indicate the
special requirement(s) for entering the laboratory.
At present these precautions are not applicable for
studies with W.
dermatitidis.
4.
Laboratory personnel will be required to receive
appropriate immunizations or tests for the agents handled or
potentially present in the laboratory (e.g., hepatitis B
vaccine or TB skin testing), should this ever become
necessary.
5.
When appropriate, considering the agent(s) handled,
baseline serum samples for laboratory and other at-risk
personnel will be collected and stored.
Additional serum specimens may be collected
periodically, depending on the agents handled or the function
of the facility. At
present these precautions are not necessary for studies with W.
dermatitidis.
6.
All personnel will be advised of these special hazards,
should they materialize and will be required to read and
follow instructions on practices and procedures as put forth
in this manual.
7.
All laboratory personnel will receive appropriate
training on the potential hazards associated with their work ,
necessary precautions to prevent exposures, and exposure
evaluation procedures. Personnel
will receive annual updates, or additional training as
necessary for procedural or policy changes.
8.
A high degree of precaution must always be taken with
any contaminated sharp items, including needles and syringes,
slides, pipettes, capillary tubes, and scalpels.
Needles and syringes or other sharp instruments should
be restricted in the laboratory for use only when there is no
alternative, such as parenteral injection, phlebotomy, or
aspiration of fluids from laboratory animals and diaphragm
bottles. Plasticware
should be substituted for glassware whenever possible.
a.
Only needle-locking syringes or disposable
syringe-needle units (i.e., needle is integral to the syringe)
should be used for injection or aspiration of infectious
materials. Used
disposable needles must not be bent, sheared, broken,
recapped, removed from disposable syringes, or otherwise
manipulated by hand before disposal; rather, they must be
carefully placed in conveniently located puncture-resistant
containers used for sharps disposal.
Non-disposable sharps must be placed in a hard-walled
container for transport to a processing area for
decontamination, preferably by autoclaving.
b.
Syringes which re-sheathe the needle, needle-less
systems, and other safe devices must be used when appropriate.
c.
Broken glassware must not be handled directly by hand,
but must be removed my mechanical means such as a brush and
dustpan, tongs, or forceps.
Containers of contaminated needles, sharp equipment,
and broken glass must be decontaminated before disposal,
according to local, state, or federal regulation.
9.
Cultures, tissues, or specimens of body fluids must be
placed in a container that prevents leakage during collection,
handling, processing, storage, transport, or shipping.
10.
Laboratory equipment and work surfaces must be
decontaminated with an appropriate disinfectant on a routine
basis, after work with infectious materials is finished, and
especially after overt spills, splashes, or other
contamination by infectious materials.
Contaminated equipment must be decontaminated according
to any local, state, or federal regulations before it is sent
for repair or maintenance or packaged for transport in
accordance with applicable local, state, or federal
regulations, before removal from the facility.
11.
Spills and accidents which could result in overt
exposures to infectious materials must be immediately reported
to PJS. Medical
evaluation, surveillance, and treatment will be provided as
appropriate and written records must be maintained.
C.
Safety Equipment
(Primary Barriers)
1.
Properly maintained biological safety cabinets,
preferably Class II, and other appropriate personal protective
equipment or physical containment devices must be used
whenever:
a.
Procedures with a potential for creating infectious
aerosols or splashes are conducted.
These may include centrifuging, grinding, blending,
vigorous shaking or mixing, sonic disruption, opening
containers or infectious materials whose internal pressures
may be different from ambient pressures, inoculating animals
intranasally, and harvesting infected tissues from animals or
eggs.
b.
High concentrations or large volumes of infectious
agents are used. Such
material may be centrifuged in the open laboratory if sealed
rotor heads or centrifuge safety cups are used, and if these
rotors or safety cups are opened only in a biological safety
cabinet.
2.
Face protection (goggles, mask, faceshield or other
splatter guards) must be used for anticipated splashes or
sprays of infectious or other hazardous materials to the face,
when the microorganisms must be manipulated outside the BSC.
3.
Protective laboratory coats, gowns, smocks, or uniforms
designated for lab use only must be worn while in the
laboratory. This
protective clothing must be removed and left in the laboratory
before leaving for non-laboratory areas (e.g., cafeteria,
library, administrative offices).
All protective clothing is then either disposed of in
the laboratory or laundered by the institution; it should
never be taken home by personnel.
4.
Gloves must be worn when handling infected animals and
when hands may contact infectious materials, contaminated
surface or equipment. Wearing
two pairs of gloves may be appropriate; if a spill or splatter
occurs, the hand will therefore be protected after the
contaminated glove is removed. Gloves must be disposed of when contaminated, removed when
work with infectious materials is completed, and must not be
worn outside the laboratory.
Disposable gloves are not washed or reused.
D.
Laboratory
Facilities (Secondary Barriers)
1.
The laboratory contains a sink for handwashing.
2.
The laboratory is designed so that it can be easily
cleaned. Rugs in
laboratories are not permitted because proper decontamination
following a spill is extremely difficult to achieve.
3.
Our bench tops are impervious to water and resistant to
acids, alkalis, organic solvents, and moderate heat.
4.
Our laboratory furniture is sturdy, and spaces between
benches, cabinets, and equipment are accessible for cleaning.
5.
Our laboratory windows should never be open, because
they are not fitted with fly screens.
6.
Equipment and methods for decontamination of infectious
or regulated laboratory wastes are available (e.g., autoclave,
chemical disinfection, incinerator, or other approved
decontamination system), in ESB or MBB, for all the activities
mentioned above.
7.
An eyewash facility is readily available in 107 (faucet
mounted).
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